PROJECT SUMMARY Acute myeloid leukemia (AML) develops as a result of the stepwise acquisition of mutations ultimately resulting in malignant transformation. Mutations that alter the epigenome occur early in disease development, while mutations that activate signaling pathways occur later. I have developed an experimental system in which mutation order can be directly manipulated in vivo. Using this system, I demonstrate that mutations in the transcription factor CCAAT enhancer binding protein alpha (CEBPA) must occur prior to mutations in Colony Stimulating Factor 3 Receptor (CSF3R) in order for leukemia to develop. In Aim 1, I will investigate whether the epigenetically similar core binding factor (CBF) AML also demonstrates order dependence with co-occurring signaling mutations. I will also establish the consequences of mutation order on the epigenetic landscape of developing hematopoietic progenitors. In Aim 2, I will evaluate the role of lysine demethylase 1 as a driver of CSF3R/CEBPA mutant AML through epigenetic profiling and loss of function studies. In Aim 3, I will identify novel therapeutic approaches that reverse the epigenetic dysfunction seen in CEBPA/CBF AML and restore the differentiation potential of AML blasts. My goal is to become a successful independent investigator and a leader in the field of leukemia genomics and targeted therapy. I will continue to conduct mentored research in the Laboratory of Dr. Brian Druker, a pioneer in the field of targeted cancer therapy and Director of the Knight Cancer Institute. I will undertake additional training in the area of cancer epigenetics and bioinformatic analysis with the guidance of my mentorship committee members Dr. Lucia Carbone and Dr. Joshi Alumkal as well as my and collaborator Dr. Hisham Mohammed. I will continue to see patients with hematologic malignancy day weekly in the outpatient setting with the remainder of my time dedicated to research. Dr. Druker and my mentorship team will assist me in obtaining my first R01 grant and in navigating the transition to running an independent research group.